The development of new, more effective analgesic drugs with high affinity for biological targets is a critical challenge. The peptide HCRG21, a TRPV1 channel blocker derived from the sea anemone Heteractis magnifica, is considered as a promising candidate for a new analgesic drug.

Aim. To investigate the genetic and expression stability of the Escherichia coli strain Origami2(K12)/pLysS/pSMT3_HCRG21, which produces the SMT3_HCRG21 fusion protein.

Materials and methods. The viability of the working seed lot (WSL) during sequential passaging was assessed by calculating the cell titer in each passage. The number of generations of WSL cells was determined based on the viable cell titer values for each passage using an appropriate equation. The expression level of the fusion protein during continuous cell passaging was calculated as the ratio of the fusion protein amount to the total cellular protein in the cell lysate. Protein content was determined using the Lowry method and electrophoresis. The size of the DNA region encoding the SMT3_HCRG21 fusion protein was determined by PCR and electrophoretic separation, and Sanger sequencing was performed to confirm its genetic identity as specified in the strain passport.

Results. During continuous passaging, the cell titer has been shown to vary from 8.3·10⁷ to 1.7·10⁹ cells/mL, and the average number of generations was 10.4 ± 2.3. Following pDNA amplification of clones from 10, 90, and 140 generations the PCR fragments of approximately 600 bp in length were obtained, which correspond to the length of the nucleotide sequence of the pSMT3_HCRG21 genetic construct (541 bp). The nucleotide sequences of the obtained PCR fragments were identical to those specified in the strain passport. It was demonstrated that the average content of the fusion protein across generations remained at 16 %, indicating a high stability of the strain productivity level.

Conclusions. The biosynthetic activity and viable cell titer of the E. coli strain Origami2(K12)/pLysS/pSMT3_HCRG21 remained stable; therefore, the strain can be used for industrial production of the analgesic peptide HCRG21.

Background. For the development of HER2-targeted CAR NK cell effectors, it is important to select an NK cells subset with high cytotoxic potential and sensitivity to genetic modification. In the context of solid tumor therapy, licensed KIR⁺ NK cells expressing the activating receptor NKG2C are of particular interest.

Aim. To obtain HER2-CAR-NK cells based on NK cell subsets with different expression of KIR and NKG2C receptors for the elimination of HER2⁺ solid tumors.

Materials and methods. NK cells from an HCMV-seropositive donor were isolated by negative magnetic separation from peripheral blood mononuclear cells isolated by density gradient centrifugation. NK cell subsets with different expression of KIR and NKG2C were obtained by cell sorting and cultured with stimulation by IL-2 and feeder cells. NK cells were modified with a gene carrying the HER2-CAR construct via retroviral transduction. The cytotoxic activity of the cultures against SKOV-3 spheroids was detected by the level of degranulating CD107a⁺ NK cells and the proportion of nonviable SytoxBlue⁺ tumor cells in the spheroid.

Results. HER2-CAR-NK effectors were obtained based on subsets with different KIR and NKG2C expression. It was shown that NKG2C-expressing NK cells, regardless of KIR expression, have greater sensitivity to retroviral transduction. Upon co-incubation of the obtained cultures of NK cell subsets with HER2⁺ SKOV-3 tumor cell spheroids, in the fraction of transduced cells bearing the HER2-CAR receptor, the proportion of degranulated CD107a⁺ cells was more than 4-fold higher than the level of degranulation in the fraction of unmodified NK cells. Also, in SKOV-3 spheroids after co-incubation with KIR⁺ NKG2C⁺ transduced NK effectors, a lower proportion of viable cells was observed compared to spheroids that were incubated with KIR⁺ NKG2C^– cultures.

Conclusion. NKG2C+ NK cells exhibited greater sensitivity to transduction. Cultures of KIR⁺NKG2C⁺ HER2-CAR-NK cells eliminated HER2⁺ target cells more efficiently in a 3D model. These properties make NK cells coexpressing KIR and NKG2C a promising basis for the CAR-NK effectors development.

Background. Previously, it has been shown in animal models that the secretome of mesenchymal stromal cells (MSCs) can restore the spermatogonial stem cell niche. In a mouse model of doxorubicin-induced toxic damage to spermatogenesis, it was shown that the biodistribution of MSC secretome following administration under the tunica albuginea of the testis occurs mainly in the interstitium, which consists predominantly of Leydig cells. However, the effect of MSC secretome on Leydig cells under doxorubicin-induced damage remains unexplored.

Aim. To study the effect of MSC secretome on Leydig cells under doxorubicin-induced damage in vitro and in vivo.

Materials and Methods. Spermatogenesis damage was modeled in C57Bl/6 mice, which received intraperitoneal injections of doxorubicin at a total dose of 10 mg/kg. Three groups of animals were studied: intact, damaged, and MSC secretome-treated. For in vitro studies, doxorubicin (at concentrations ranging from 0.15 to 1.5 µM) and MSC secretome were added to Leydig cell cultures.

Results. Following doxorubicin-induced damage in vivo, mouse testis weight decreased and did not recover after MSC secretome injection. The interstitial area increased upon damage, while administration of MSC secretome tended to reduce (normalize) the interstitial area. At the same time, the number of Leydig cells did not change significantly. In in vitro studies, exposure of Leydig cells to doxorubicin induced dose-dependent cell death. Addition of MSC secretome to Leydig cells under damage conditions enhanced cell survival and testosterone secretion, resulting in an increase in total testosterone content in the Leydig cell culture medium.

Conclusion. MSC secretome can act on Leydig cells under doxorubicin-induced damage by increasing their number and secretory activity, which may contribute to interstitial recovery. These findings provide a rationale for further investigation of MSC secretome as a promising therapeutic agent for restoring testicular function.

Background. Cardiac surgery and cardiopulmonary bypass (CPB) cause ischemia-reperfusion injury, leading to endothelial dysfunction, systemic inflammation and, in some cases, multiple organ failure (MOF), which worsens the prognosis.

Aim. To study the effect of nitric oxide (NO) and molecular hydrogen (H₂) on endothelial dysfunction, myocardial contractility during surgery, and postoperative multiple organ failure.

Materials and methods. The study included 123 patients randomized into 4 groups: 1^st (n = 30) – control; 2^nd (n = 33) – isolated supply of NO (40 ppm) in the CPB circuit; 3^rd (n = 30) – isolated supply of H2 (2 vol%) into the CPB circuit; 4th (n = 30) – combined supply of NO (40 ppm) and H₂ (2 vol%) into the CPB circuit. During surgery, the functional parameters of the heart were studied using transesophageal echocardiography; the state of endothelial cells – by analysis of circulating endothelial cells (CECs); after surgery – biochemical blood parameters and clinical outcomes.

Results. The action of NO and H₂ improved the contractile function of the myocardium, reduced the increase in AST and ALT activity, reduced the urea content compared with the control, maintaining bilirubin (with the combined action of gases) and creatinine within the initial values, which indicates a multi-organ protective effect. Against the background of the action of NO and H₂, a more pronounced decrease in circulating endothelial cells was noted, associated with a lower incidence of postoperative complications and MOF. The combined use of NO and H₂ caused a multi-organ protective effect superior to the effect of each gas.

Conclusion. Extracorporeal administration of NO and H₂ is an effective strategy for correcting myocardial and endothelial dysfunction, which helps prevent the development of multiple organ failure in cardiac surgery patients.

Background. Induction of labor (IOL) is a widely used obstetric intervention. However, the rate of cesarean section (CS) following IOL may exceed 50% in certain subgroups. Existing predictive models predominantly rely on antenatal factors, fail to account for nonlinear relationships, and do not always demonstrate sufficient discriminative ability.

Aim. Development, validation, and interpretation of a predictive model for predicting the outcome of IOL in term pregnancy using machine learning methods.

Materials and methods. This retrospective cohort study, based on a hospital registry (June 2024 – June 2025), included 1,670 women with singleton pregnancies with gestational age of ≥37 weeks who underwent IOL (amniotomy, oxytocin). Outcomes were defined as successful IOL (vaginal delivery) and unsuccessful IOL (CS). For preliminary feature selection, feature importance was calculated using logistic regression, logistic regression with L1 regularization (Lasso), and random forest to eliminate non-informative variables. The final predictive model was built using the random forest algorithm. SHAP analysis was used to interpret the influence of the selected features on the model predictions.

Results. The CS rate was 21.1% (n=352). Seven features were included in the final model (in descending order of importance): the time interval from IOL initiation to the onset of regular uterine contractions, parity, estimated fetal weight by ultrasound, need for pre-induction of labor, maternal age, maternal body mass index, and gestational age. The model achieved an AUC-ROC of 0.904 (95% CI 0.846–0.951) on the test set, with a sensitivity of 75%, specificity of 92%, and net benefit of 0.184. SHAP analysis enabled interpretation of individual predictions.

Conclusion. The developed random forest-based model, which incorporates an intrapartum predictor (time interval to regular contractions), demonstrates high predictive performance, potential for external validation, and clinical utility, thereby supporting timely and informed decision-making regarding labor management.

Background. The use of prophylactic HPV vaccines has demonstrated effectiveness in reducing the incidence of cervical cancer. The yeast Kluyveromyces lactis is a promising platform to produce recombinant vaccine antigens, owing to its GRAS status, eukaryotic post translational modification capabilities, and the low cost nutrient media.

Aim. To develop a producer strain based on Kluyveromyces lactis for the production of recombinant human papillomavirus type 18 L1 protein.

Materials and Methods. For the expression of the HPV18 L1 gene, two integrative vectors were constructed. The first, pKB-HPV18, with a constitutive GAP promoter, and the second, pKB-HPV18, with an inducible LAC4 promoter. K. lactis cells were transformed with the obtained vectors by electroporation. Recombinant strains were cultivated in YEPm medium. Protein synthesis induction for the K. lactis/pKB-HPV18 strain was performed using lactose. The target protein was isolated from cell lysates by ion exchange chromatography (SP Seplife). Protein identification was carried out using liquid chromatography–tandem mass spectrometry (LC MS/MS). All experiments were performed in three biological replicates. Statistical significance was assessed using Student’s t-test (p < 0.05).

Results. Producer strains were obtained for both constructs. The biomass yield was 20 g/L for K. lactis/pKB-HPV18 and 34 g/L for K. lactis/pKB-HPV18. The purity of the target protein after a single ion-exchange chromatography step reached 70–80% for the constitutive system and approached 90% for the inducible system. The identity and structural integrity of the obtained protein were confirmed using LC MS/MS.

Conclusion. Two K. lactis producer strains capable of synthesizing HPV18 L1 protein were successfully constructed. The system with the inducible LAC4 promoter showed higher biomass yield and target protein purity compared to the constitutive system. These results provide a foundation for the further development of an affordable production system for the HPV18 vaccine antigen.

The review presents an analysis of current literature data on cardiac sarcoidosis (CS), the frequency of which varies widely in the structure of systemic sarcoidosis – from 3 to 39%. Many authors have noted a general upward trend in the prevalence and incidence of both systemic and CS, with variations depending on geographical region and ethnicity. It is suggested that the clinical phenotypes of CS differ depending on ethnic predisposition and sex. The role of human leukocyte antigen (HLA) polymorphism and specific alleles in the development of cardiac involvement in sarcoidosis has been investigated. The possible influence of infectious agents (Cutibacterium acnes, Mycobacterium tuberculosis, etc.) as an initiating factor in the development of the disease is considered. Non-caseating epithelioid granulomas, a specific morphological feature of sarcoidosis, can be localized in any part of the myocardium, leading to rhythm and conduction disturbances, the development of ventricular aneurysms, and heart failure. Multinucleated giant cells are detected in non-caseating epithelioid granulomas in no more than 20% of CS cases, and the absence of this specific pathomorphological feature does not rule out cardiac involvement in sarcoidosis. The staging of granulomatous inflammation supports the use of surrogate histopathological signs, including microgranulomas, clusters of M1-macrophages, signs of lymphangiogenesis, confluent fibrosis and fatty infiltration of the myocardium.

Background. Malaria is a global public health problem. Traditional blood microscopy, the gold standard of diagnosis, is associated with high labor burden, personnel shortages in endemic regions, and subjective assessment. Automation of not only detection but also determination of the pathogen’s developmental stage is critically important for prescribing stage-specific therapy.

Aim. To develop and evaluate the effectiveness of a deep computer vision model for classifying four key stages of the erythrocytic cycle of Plasmodium falciparum based on microphotographs.

Materials and methods. A convolutional neural network with the ResNet-18 architecture was used as the basis. Training was conducted using transfer learning on the PfMorphoBank database, assembled by the authors. The images were divided into four target classes: young trophozoites, mature trophozoites, schizonts, and gametocytes. Classification quality was assessed on validation and held-out test sets using accuracy, macro-precision, macro-recall, and macro-F1.

Results. On the held-out test set, the fine-tuned ResNet-18 achieved 80.83% accuracy; macro-precision was 0.812, macro-recall was 0.808, and macro-F1 was 0.806. Error analysis showed better recognition of mature trophozoites and the greatest difficulty with gametocytes and morphologically similar late-stage forms.

Conclusion. The use of deep neural networks demonstrates high potential in solving the problem of differential diagnosis of malaria parasite stages. The model showed acceptable quality metrics, proving the fundamental feasibility of creating automated prompting systems.

Free radical homeostasis is a state of a biological system characterized by a balance between the production of free radicals and their neutralization by the body’s antioxidant defense systems. A disruption of this balance leads to a change in the redox state of the cell, which contributes to the development of oxidative stress, a key link in the pathogenesis of many diseases. Considering that most current studies assess the parameters of the redox system in venous blood samples, the collection procedure of which is invasive and, accordingly, has a number of limitations, there is a need to search for alternative biological matrices suitable for the analysis of redox parameters.

Aim. To analyze current concepts regarding the potential use of tear fluid, saliva, and urine as promising biological matrices for assessing the state of free radical homeostasis. This review examines the characteristics of the formation and composition of tear fluid, saliva, and urine, as well as the sources of free radicals, products of oxidative modification of biomolecules, and components of the antioxidant defense system detected in them. The factors influencing the interpretation of free radical homeostasis parameters in these biological matrices are analyzed, including formation specifics, sample collection methods, the impact of local and systemic pathological processes, approaches to data normalization, and external factors. It is shown that tear fluid provides high diagnostic value for assessing local changes occurring on the ocular surface and in the anterior segment of the eye. Saliva reflects a combination of local and systemic changes, while urine is the most extensively studied and standardized biological matrix, with parameters primarily reflecting established systemic alterations in free radical homeostasis.

Thus, tear fluid, saliva, and urine possess varying diagnostic utility for assessing local and systemic changes in free radical homeostasis. Future prospects in this field are associated with the development of standardized approaches to the analysis of these biological matrices and a comprehensive assessment of free radical homeostasis.

Background. Ethoxylated nonylphenol (ENP) is a surfactant characterized by high toxicity and endocrine-disrupting activity that persists after partial biodegradation. In nanobioremediation, sublethal concentrations of nanoparticles can induce a hormetic effect, stimulating biofilm formation and enhancing the catabolic potential of microorganisms. Copper oxide nanoparticles (CuO NPs) at sublethal concentrations modify cell surface properties and stimulate biofilm formation, thereby enhancing the biocatalytic potential of bacteria of the genus Rhodococcus in bioremediation processes. However, the mechanisms of Rhodococcus biofilm adaptation to organic xenobiotics and nanometals remain poorly understood.

Aim. To evaluate the effect of combined exposure to ENP and sublethal concentrations of CuO NPs on the morphology, surface properties, and biofilm formation of Rhodococcus ruber.

Materials and methods. Strains R. ruber IEGM 1263 and IEGM 71 from the Regional Specialized Collection of Alkanotrophic Microorganisms were used. The minimum inhibitory concentration (MIC) was determined under exposure to ENP (0.16–100 g/L) and CuO NPs (0.0007–0.5 g/L). Biofilms were cultivated on polypropylene discs. The ζ-potential of cells was measured by dynamic light scattering, while cell viability and surface morphology were analyzed using combined confocal laser scanning and atomic force microscopy (CLSM-AFM).

Results. Combined exposure to ENP and CuO NPs reduced rhodococcal viability by 20–80 %, whereas the overall toxic effect was mainly determined by copper oxide nanoparticles. Biofilms of the resistant strain R. ruber IEGM 1263 grown in the presence of sublethal concentrations of ecotoxicants (1–10 g/L ENP and 0.001 g/L CuO NPs) were characterized by decreased adhesion to the carrier (2.6-fold), cell coaggregation, a more negative ζ-potential (−34.2 mV), and increased cell surface roughness (up to 4.8 nm) compared with the control.

Conclusion. Morphophysiological adaptations of Rhodococcus ruber cells promoting biofilm formation and increasing bacterial resistance to complex chemical stress were identified. The obtained data confirm the potential of using Rhodococcus biofilms for bioremediation of ecosystems contaminated with organic xenobiotics and nanometals.

Background. Lactobacilli, being representatives of the normobiota, are of great interest as the most important and promising biotechnological targets for the development of probiotics and functional nutrition. The key criterion for strain selection is proven safety, specifically the absence of virulence and antimicrobial resistance (AMR) genes. An important aspect of the biosafety of lactobacilli is the absence of plasmids in the genome.

Aim. To evaluate the safety of lactobacilli strains based on their phenotypic susceptibility to antimicrobials (AMs) and whole-genome analysis for the evidence-based selection of candidate strains for probiotic consortia.

Materials and methods. The study included 34 strains of lactobacilli isolated from the intestinal biotope of healthy and apparently healthy children and adolescents in the Irkutsk region. The strains were identified using MALDI-TOF mass spectrometry and whole-genome sequencing. Susceptibility to AMs was determined using the disk diffusion method. The genomes of lactobacillus strains were screened for the presence of AMR, virulence, and plasmid sequences.

Results. A phenotypic analysis of the susceptibility of lactobacilli strains to AMs revealed both collection-wide characteristics and species-specific features, mainly in relation to oxacillin and penicillin G. The genes for AMR and virulence were absent in the genomes of all analyzed lactobacillus strains. Plasmids were found in the genomes of 26 of the 34 strains. In terms of horizontal transfer, 37 plasmids were non-mobilizable, 22 were mobilizable, and 1 was conjugative. Strains with mobile and conjugative plasmids were excluded from the list of candidate strains for probiotic consortia.

Conclusion. The proposed algorithm for selecting lactobacilli strains, including the assessment of phenotypic susceptibility to AMs, genome screening for the presence of AMR and virulence genes, and mobile plasmid elements, enables the selection of safe strains for the further development of probiotic consortia.

Background. Numerous studies link olfactory impairment to accelerated cognitive decline in neurodegenerative diseases, yet the mechanism by which the olfactory bulb participates in these processes remains unclear. βIII-tubulin is one of the most commonly used neuronal markers, but data on its distribution in the olfactory bulb are limited.

Aim. To analyze the distribution of βIII-tubulin in the olfactory bulbs of men and women during aging.

Methods. The study was performed on autopsy material. Olfactory bulbs were obtained from 10 men and 14 women of elderly (60–74 years, n = 11) and senile (75–89 years n = 13) ages. Histological sections were stained with hematoxylin and eosin, Nissl stain, and Mallory’s trichrome for cytoarchitectonic analysis. Immunohistochemical analysis was performed using primary antibodies against neuron-specific βIII-tubulin. Morphometric and statistical analyses were carried out.

Results. βIII-tubulin immunoreactivity was mainly detected in the olfactory nerve fiber layer and the glomerular layer. Analysis of sex differences revealed a significant (p = 0.006) predominance of the βIII-tubulin reaction area in olfactory bulb sections obtained from women. This aligns with research findings showing that women have nearly twice as many neurons in their olfactory bulbs and typically exhibit better odor identification ability compared to men. Despite a trend towards a decrease in βIII-tubulin levels with age, no statistically significant differences were found.

Conclusion. The study revealed sexand age-related variability in the distribution of βIII-tubulin in the human olfactory bulb. Therefore, these factors should be taken into account when performing functional olfactory diagnostics.

Introduction. Predicting the outcomes of traumatic brain injury (TBI) depending on the degree of cognitive impairment is important for optimizing diagnostic and therapeutic care, but cognitive impairments are rarely used in predictive models.

Aim. To develop models for predicting the outcomes of mild and moderate TBI in elderly patients, taking into account cognitive status.

Material and methods. A total of 125 patients aged 60–74 years with mild and moderate TBI were examined on days 8–9 and 20–21, respectively. TBI outcomes were classified into 3 groups: 1 – patients’ ability to perform motor and sensory actions, 2 – mood and interpersonal interaction, 3 – social contacts and work. Cognitive impairments were detected using the Mini-Mental State Examination (MMSE). The Statistica 17.0 program and regression analysis were used to develop predictive models of the above-mentioned TBI outcomes.

Results. Weak inverse correlations between mild TBI severity and TBI outcomes were established. However, the association of moderate TBI severity with outcomes was higher than for mild TBI, especially for participation in social contacts and work (r = -0.452; p < 0.01) and for mood and interpersonal interaction (r = -0.406; p < 0.01). In all cases, the outcomes of TBI with cognitive impairments had a moderate positive correlation, with correlation coefficients from 0.339 to 0.372 (p < 0.01). To predict the ability of patients to perform motor and sensory actions (y₄), taking into account the score of cognitive impairment (x₄), an adjusted model y₄ = 7,389–1,724x4 was created. To predict mood and interpersonal interaction, an adjusted model y₅ = 8,241–1,406x5; to predict patient participation in social contacts and work, an adjusted model y₆ = 7,895– 1,469x6, where x₅ and x₆ are the scores of cognitive impairments on the MMSE scale.

Conclusion. The results show the negative impact of cognitive impairment on the outcomes of TBI, for the prediction of which prognostic models have been developed.

Post-COVID syndrome (PCS) is accompanied by neurological manifestations. The routes of virus penetration and the pathogenetic mechanisms triggered by COVID-19 in the infected body play an important role.

Aim. To provide a clinical and neurophysiological description of the neurological manifestations of post-COVID syndrome.

Methods. A total of 225 women with PCS were examined. Clinical, neurophysiological (electroencephalography, electroneuromyography), and neuropsychological examinations were performed.

Results. The main clinical syndrome was asthenia (100 % of cases); cognitive and emotional-volitional disorders were recorded (70.8 % and 36.2 %, respectively). Hyposmia was observed (55 %), distorted perception of odors (parosmia) was recorded in 20 % of cases, hyperosmia and anosmia – in 5 %, and 15 % of the examined patients had no impairment of smell. In patients with PCS, distal polyneuropathy was diagnosed in 11.1 % of cases, confirmed by electroneuromyography. According to the electroencephalogram (EEG), a change in coherence in the alpha-band was revealed. An increase in interhemispheric connections in the temporal regions was observed, and coherence in the frontal areas decreased compared to normal values. Neuropsychological testing showed a decrease in cognitive functions, manifested in changes in neuropsychological indicators reflecting impairments in short-term (auditory-verbal) and long-term memory, and reciprocal coordination.

Conclusion. The main clinical manifestations of PCS in women are asthenia of varying severity, cognitive and emotional-volitional disorders with an anxiety-depressive component, disorders of sensory functions (smell and taste), and polyneuropathy of the extremities. The study revealed impaired indicators reflecting the state of shortterm (auditory-verbal) and long-term memory, as well as reciprocal coordination.

Objective assessment of postoperative eye refraction in patients with implanted intraocular lenses (IOLs) featuring addition zones or a modified wavefront profile is challenging using conventional methods.

Aim. To analyze the diagnostic value of the method of objective refractive topographic mapping of the eye in patients with IOLs of complex optical design.

Material and Methods. A total of 198 patients (mean age 66.4 ± 7.8 years) with various types of IOLs – monofocal, multifocal, and extended depth of focus lenses were initially screened. Inclusion criteria: the absence of pathology of the cornea, vitreous, retina, or optic nerve; a postoperative follow-up period of six months or more. Exclusion criteria: uncorrected distance visual acuity of 0.7 or worse, opacity or folds of the posterior lens capsule. After applying inclusion and exclusion criteria and performing random eye selection (right or left) for eligible participants, 192 eyes were included in the final analysis. One eye per subject was chosen at random to avoid inter-eye correlation. All patients underwent a comprehensive ophthalmological examination, including objective assessment, visometry, tonometry, depth of focus (DoF) calculation based on the defocus curve (the diopter range within which visual acuity remains no lower than 0.5), wavefront aberrometry (Peramis, SCHWIND eye-tech-solutions GmbH, Germany) with refractive topographic map (RTM) construction and calculation of the refractive inhomogeneity index.

Results. The construction of RTMs revealed the distribution of refractive indicators across the entire scanning area and the presence of distinct addition zones in eyes with multifocal IOLs and extended depth of focus IOLs. Significant differences were found in the refractive inhomogeneity index values between groups (H(7) = 46.24, p = 0.0001) and varying levels of its consistency with DoF.

Conclusion. The method of pyramidal sensor-based aberrometry with refractive topographic mapping represents a highly informative approach for the objective assessment of the eye’s optical system in patients with complex-design IOLs and could serve as a foundation for the development of personalized cataract surgery.

Introduction. Obesity in adolescents is a global medical and social problem associated with a progressive increase in incidence. The role of structural changes in the colonic microbial community in the pathogenesis of exogenous-constitutional obesity in adolescents remains insufficiently studied, and data on characteristic taxonomic alterations are contradictory.

Aim. To study the characteristics of the qualitative and quantitative composition of the colonic microbiota in adolescents with obesity and normal body weight.

Materials and methods. A cross-sectional comparative study was conducted. The main group consisted of 20 adolescents aged 12–17 years with obesity (BMI SDS ≥ +2.0), and the control group consisted of 20 adolescents with normal body weight. The taxonomic composition of the microbiota was examined by real-time PCR using the “Colonoflor-16” test system. Statistical analysis was performed using the Mann – Whitney and Fisher tests.

Results. Compared to the control group, changes in the taxonomic composition of the microbiota in adolescents with obesity were characterized by a significant decrease in total bacterial mass and the abundance of key symbionts, including Bifidobacterium spp. A decrease in the level of Lactobacillus spp. was observed in both study groups without statistically significant intergroup differences. A deficiency of butyrate producers and key participants in fiber fermentation was identified, including Faecalibacterium prausnitzii, Eubacterium rectale, and Akkermansia muciniphila, as well as a trend toward a decrease in Roseburia inulinivorans, Ruminococcus spp., Prevotella spp., and Blautia spp. Concurrently, an increase was recorded in the abundance of Bacteroides spp., opportunistic pathogens (Enterobacter spp., Escherichia coli, Streptococcus spp., Enterococcus spp.), and the Bacteroides spp. / Faecalibacterium prausnitzii ratio. In the main group of adolescents, Clostridium difficile, Clostridium perfringens, Staphylococcus aureus, Acinetobacter spp., and fungi of the genus Candida were detected.

Conclusion. Adolescents with obesity exhibit a specific pro-inflammatory profile of the colonic microbiota, characterized by depletion of symbionts, deficiency of butyrate producers, and overgrowth of opportunistic taxa. The obtained data identify potential targets for personalized microbiome correction in the comprehensive therapy of obesity.

Background. The Irkutsk Region remains a region with a high burden of multidrug-resistant tuberculosis and a challenging epidemiological situation regarding HIV infection. For immunocompromised individuals, the spread of nontuberculous mycobacteria (NTMB) poses a significant threat, as they cause pulmonary mycobacteriosis – a condition that mimics tuberculosis but requires differential diagnosis and distinct treatment approaches. Regional studies on NTMB prevalence remain limited. Given the low resolution of standard microbiological techniques, the search for molecular targets for NTMB species identification is essential.

Aim. To evaluate the species diversity of NTMB isolated from patients with suspected pulmonary tuberculosis and mycobacteriosis in the Irkutsk Region.

Materials and methods. A retrospective analysis was performed on NTMB strains obtained from 63 patients treated at the Irkutsk Regional Clinical Tuberculosis Hospital between 2018 and 2024. Species identification was performed for 50 isolates using sequencing of the 23S rRNA and groEL2 genes fragments, followed by phylogenetic analysis.

Results. Representatives of the Mycobacterium avium complex (MAC) dominate among the studied strains (65.9%), including M. avium (53.7%) and M. intracellulare (12.2%). Joint analysis of the 23S rRNA and groEL2 gene fragments ensured the differentiation of species within the MAC, as well as the identification of a wide range of slowly and rapidly growing NTMB (M. paragordonae, M. abscessus, M. fortuitum, M. holsaticum, and M. farcinogenes/M. senegalense) and non-mycobacterial pathogens (Nocardia farcinica, Streptococcus oralis, Staphylococcus epidermidis, Actinomadura sp.).

Conclusion. The taxonomic spectrum of NTMB isolated in the Irkutsk Region was characterized over time. Sequencing of the 23S rRNA and groEL2 genes allows overcoming the challenge of NTMB identification.

Background. It is an undeniable fact that colon polyps are associated with colorectal cancer. The main measure for preventing colorectal cancer is the removal of epithelial neoplasms. However, the effectiveness of resection depends on various factors.

Aim. To determine whether the size, location, macroscopic type, morphology, and the method of resection influence the effectiveness of removal of laterally spreading tumors in the colon and rectum.

Methods. We conducted a retrospective analysis of cases of removal of LST type epithelial neoplasms in the colon and rectum in 2023. Endoscopic resection was performed in 165 patients with benign epithelial neoplasms ranging in size from 11 to 30 mm, with an average size of 15 ± 4.43 mm. Eight patients with giant epithelial neoplasms measuring 6–14 cm in length, up to 2/3 of the circumference of the rectal lumen, underwent SILS-transanal resection. Patients were invited for follow-up examination at 3, 6 and 12 months after removal.

Results. We found a relationship between the relapse of the LST type epithelial neoplasm and the size (from 20 to 30 mm, p = 0.010), macroscopic type (granular mixed-nodular, p = 0.012) and morphology (tubulovillous adenoma, p = 0.028) of the previously resected lesion. In all cases, when removing the epithelial lesion, the edges of the wound on the mucosa were carefully examined for residual tissue. If found, the residual tissue was removed completely. We suggest that tubulovillous adenoma cells might have been implanted into the submucosa with an injector or a snare during removal of the lesion. The SILS-TAR group showed no relapses. However, the SILS-port (COVIDIEN) was recently discontinued.

Conclusion. The possibility of accidental implantation of benign adenoma cells into the submucosa with an injector or a snare during resection warrants further study. It is also necessary to continue the search for and development of new affordable and effective methods for removing giant villous adenomas of the rectum.

Background. Squalene represents a promising basis for the development of novel adjuvant compositions with efficacy comparable to traditional adjuvants but with an improved safety profile.

Objective. To develop a novel squalene based adjuvant composition and evaluate its ability to enhance the humoral immune response against three different viral antigens — the receptor binding domain (RBDd) of the SARS CoV 2 Delta variant S protein, domain III (WDIII) of the West Nile virus E surface protein, and the surface glycoprotein (trGPM) of the Marburg virus.

Materials and methods. The adjuvant was prepared as an oil-in-water emulsion containing squalene, lecithin and Tween 80, followed by ultrasonic homogenization. BALB/c mice were immunized with two intramuscular injections 14 days apart. Specific IgG antibody titers were determined by ELISA. Statistical analysis was performed using the Mann–Whitney U-test.

Results. The ASV24 emulsion remained stable for 12 months. When immunized with the WDIII antigen produced in the prokaryotic E. coli system, the adjuvant activity of ASV24 was comparable to that of aluminum hydroxide. For the RBDd antigen produced in Chinese hamster ovary (CHO) cells, a trend toward higher titers was observed in the ASV24 group (3.6 × 10⁶) compared to aluminum hydroxide (9.6 × 10⁵), although no statistically significant differences were found. A pronounced advantage of ASV24 was demonstrated for trGPM, also produced in CHO cells: the mean reciprocal titer was 1,5 orders of magnitude higher than that in the aluminum hydroxide group.

Conclusion. The ASV24 adjuvant composition is stable, and its efficacy can surpass that of aluminum hydroxide when using complex multimeric antigens produced in eukaryotic expression systems. These findings suggest that ASV24 is a promising adjuvant for the development of subunit vaccines.

Background. We previously developed a protocol for generating macrophages with pronounced anti-inflammatory and reparative/neuroregenerative activity, polarized by efferocytosis (M2(Eff)). Due to their unique secretome, these cells are promising candidates for cell therapy of neuroinflammatory diseases. However, the expression of the inhibitory receptors PD-1 and TIM-3 on M2(Eff) and its relationship with cytokine production remain unclear.

Aim. To characterize PD-1 and TIM-3 expression on M2(Eff) macrophages and determine their role in the regulation of proand anti-inflammatory cytokine production (IL-10 and TNF-α).

Materials and methods. Macrophages were differentiated from peripheral blood monocytes of healthy donors. PD-1 and TIM-3 expression, as well as IL-10 and TNF-α production, were assessed by flow cytometry. To evaluate the contribution of PD-1 and TIM-3 to cytokine secretion, cells were pre-treated with blocking antibodies, followed by assessment of intracellular IL-10 and TNF-α levels.

Results. M2(Eff) exhibited higher PD-1 and TIM-3 levels compared to M1 cultures, primarily due to cells co-expressing both markers. M2(Eff) also had higher levels of PD-1 and lower levels of TIM-3 compared to “classical” M2 macrophages polarized by IL-4. The proportion of IL-10-positive cells was higher in M2(Eff) cultures expressing PD-1, as well as in TIM-3-expressing cells after lipopolysaccharide stimulation, compared to the negative populations. TNF-α levels were higher in TIM-3-positive macrophages compared to TIM-3-negative ones. PD-1 blockade during lipopolysaccharide stimulation increased the proportion of TNF-α-expressing cells, whereas TIM-3 blockade had no such effect.

Conclusion. M2(Eff) macrophages induced by apoptotic cells display a unique PD-1 and TIM-3 expression profile. Moreover, PD-1 and TIM-3 are associated with IL-10 and TNF-α production.

Background. The clinical potential of magnesium implants depends on their degradation behavior and the response of surrounding tissues. For Mg–2Zn–2Ga, the correspondence between radiological and morphological changes in a paired subcutaneous model with a titanium control has not been adequately studied.

Aim. To compare MSCT signs of Mg–2Zn–2Ga degradation and gas formation with the morphological response of soft tissues relative to titanium at 1–11 weeks.

Materials and methods. The study was performed on 20 male Wistar rats. Discs measuring 7 mm in diameter and 4 mm in thickness were implanted subcutaneously over the scapulae of each animal: Mg–2Zn–2Ga on the left and titanium on the right. At weeks 1, 2, 3, 4, and 11 (four animals per time point), MSCT, macroscopic assessment, and histological examination of both implantation sites were performed.

Results. At week 1, a gas cavity was detected around Mg–2Zn–2Ga, and a slight reduction in radiodensity was observed. At weeks 2–4, radiological changes became more apparent; by week 4, the magnesium specimen was visually smaller than the stable titanium implant. At week 11, a residual gas cavity and reduced radiodensity and dimensions of Mg–2Zn–2Ga persisted. Histologically, the early cavity with a thin fibrous capsule and lymphohistiocytic infiltration was followed by a cellular rim and multiple cavities with fragmentation of adjacent muscle fibers; occasional foreign-body giant cells were detected by week 11. Orderly connective-tissue organization predominated around titanium. Crepitus over the magnesium implant was observed in 3 of 20 animals (15 %).

Conclusion. Comparison of MSCT and histology revealed a concordant time course of Mg–2Zn–2Ga degradation signs and surrounding tissue remodeling that differed from the response to titanium. Early changes developed at weeks 2–4 and persisted at week 11. These time points and features may be used in subsequent quantitative preclinical assessment.

Background. Gestational diabetes mellitus (GDM) is a serious and widespread health problem. Analysis of existing animal models of GDM induced by a high-fat diet (HFD) and administration of streptozotocin, which destroys pancreatic β-cells, reveals significant differences in dosage and duration of drug administration.

Aim. Optimization of the protocol for administering streptozotocin to female C57BL/6 mice to model GDM accompanied by persistent hyperglycemia and impaired glucose tolerance during pregnancy for testing drugs.

Materials and Methods. Female C57BL/6 mice were kept on a HFD throughout the experiment. Before pregnancy, the mice were injected three or four times with STZ at a dose of 60 mg/kg. Body weight, feed and water intake, and blood glucose levels in the females were assessed, glucose tolerance test and pathomorphological analysis of the pancreas were performed, and the reproductive capacity of dams and survival of the offspring were assessed.

Results. Four consecutive injections of streptozotocin at a dose of 60 mg/kg contribute to the development of hyperglycemia and lead to an increase in blood glucose concentration in females in a glucose tolerance test during pregnancy and lactation. Animals exhibit more pronounced polydipsia, atrophic changes in the pancreatic endocrine system, as well as a tendency to reduce litter size and offspring survival. The use of three streptozotocin doses causes hyperglycemia, leading to an increase in the area under the curve in the glucose tolerance test only during pregnancy.

Conclusion. Three injections of streptozotocin at a dose of 60 mg/kg are sufficient to induce hyperglycemia and impaired glucose utilization in pregnant C57BL/6 mice fed a HFD. This administration regimen causes temporary metabolic disturbances and does not affect the reproductive function of animals, making it suitable for use as a model to assess the efficacy of hypoglycemic drugs that could be used in GDM treatment.

Endovascular embolization coils are a key tool for minimally invasive vessel occlusion; however, their widespread use in Russian practice is limited by high cost and dependence on imported supplies. The development of Russian analogues requires a comprehensive assessment of the biocompatibility of designs that combine a metal frame with polymer fibers.

Aim. To evaluate the biocompatibility of experimental prototypes of embolization coils made of stainless steel with polymer fibers (silk, poly(glycolide-co-lactide), polypropylene, polyethylene terephthalate, polyvinyl chloride with barium sulfate).

Materials and Methods. Coil samples were fabricated by mechanical winding of stainless steel wire followed by manual fixation of polymer fibers in the inter-coil spaces. Hemolytic activity was assessed by the degree of hemolysis of human red blood cells upon contact with sample extracts. Cytotoxicity of the experimental coil samples with fibers was evaluated using human fibroblasts of the FEC-16-2 line by direct (co-culture) and indirect (cultivation in material extracts) methods.

Results. All studied samples met standard biocompatibility criteria: the hemolysis level did not exceed acceptable values (less than 2 %), and no cytotoxic effect was detected. The lowest hemolysis values were recorded for coils with silk fibers (0.005 ± 0.004 %). Samples with poly(glycolide-co-lactide) showed a somewhat higher hemolysis level (1.184 ± 0.212 %), which may be attributed to hydrolytic degradation of the polymer with a local decrease in pH.

Conclusion. Silk, polypropylene, polyethylene terephthalate, and radiopaque fibers based on polyvinyl chloride with barium sulfate are promising materials for the development of Russian embolization coils, offering a combination of hemocompatibility and functionality. The use of poly(glycolide-co-lactide) requires additional evaluation of long-term safety. The obtained results justify further preclinical studies, including the assessment of thrombogenicity and stability of fiber fixation under simulated blood flow conditions.

Introduction. Serine proteases are the most numerous catalytic type of proteolytic enzymes and perform a variety of functions in the body. They participate in digestion, act as blood clotting factors, are components of complement and the kallikrein-kinin system, among others. Trypsin is the most studied serine protease. It is known that in certain pathological processes and conditions, such as pancreatitis and pancreatic necrosis, trypsin enters the systemic circulation. This can lead to the development of thrombosis and thromboembolism, but the mechanisms underlying these disorders remain poorly understood. One possible mechanism may be trypsin-mediated platelet activation followed by aggregation.

Aim. To investigate the ability of trypsin to induce platelet aggregation in vitro. Methods. Experiments were conducted on platelet-rich plasma obtained from 22 relatively healthy volunteers. Platelet aggregation by trypsin at final concentrations of 0.5, 1, 2.5, 5, and 10 μM was studied using optical aggregometry. Trypsin-induced platelet aggregation was then recorded in prothrombin-deficient plasma.

Results. Trypsin is capable of inducing platelet aggregation in vitro. The minimal effect was observed with the addition of 2.5 μM. Addition of 5 and 10 μM trypsin to platelet plasma induced pronounced and irreversible platelet aggregation. Trypsin-induced aggregation occurs in both heparinized plasma and plasma deficient in prothrombin complex factors.

Conclusion. The study revealed proaggregatory effects of trypsin that are thrombin-independent and can occur without its participation.